GSM6809657: B. fragilis - deoxycholate 20 min - rep 2; Bacteroides fragilis; RNA-Seq
After treatment, culture was pelleted at 15,000 x g for 1 minute, supernatant was removed, and pellets were resuspended in 1 ml TRIzol and stored at -80°C. Samples were heated at 65°C for 10 min. Then, 200 µl chloroform was added, samples were vortexed, and incubated at room temperature for 5 min. Samples were centrifuged at 17,000 x g for 15 min at 4°C, then the upper aqueous phase was transferred to a fresh tube, an equal volume of 100% isopropanol was added, and samples were stored at -80°C overnight. Samples were centrifuged at 17,000 x g for 30 min at 4°C, the supernatant was removed, and the pellets were washed with cold 70% ethanol. Pellets were centrifuged again at 17,000 x g for 5 min at 4°C, supernatant was removed, and the pellet was allowed to dry before resuspension in 100 µl RNase-free water and incubation at 60°C for 10 min. Samples were treated with TURBO DNase and cleaned with RNeasy Mini Kit  Library preparation was performed using Illumina's Stranded Total RNA Prep Ligation with Ribo-Zero Plus kit and 10bp IDT for Illumina indices 